DOCSLIB.ORG

Characterization of the Gene Expression Profile of Human Bocavirus

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterization of the Gene Expression Profile of Human Bocavirus View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Virology 403 (2010) 145–154 Contents lists available at ScienceDirect Virology journal homepage: www.elsevier.com/locate/yviro Characterization of the gene expression profile of human bocavirus Aaron Yun Chen a,1, Fang Cheng a,1, Sai Lou a,b, Yong Luo a, Zhengwen Liu b, Eric Delwart c, David Pintel d, Jianming Qiu a,⁎ a Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS, USA b Department of Infectious Diseases, The First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, China c Blood Systems Research Institute, San Francisco, CA, USA d Life Sciences Center, University of Missouri-Columbia, Columbia, MO, USA article info abstract Article history: We have generated a quantitative transcription profile of human bocavirus type 1 (HBoV1) by transfecting a Received 16 February 2010 nearly full-length clone in human lung epithelial A549 cells as well as in a replication competent system in Returned to author for revision 9 April 2010 293 cells. The overall transcription profile of HBoV1 is similar to that of two other members of genus Accepted 15 April 2010 Bocavirus, minute virus of canines and bovine parvovirus 1. In particular, a spliced NS1-transcript that was Available online 10 May 2010 not recognized previously expressed the large non-structural protein NS1 at approximately 100 kDa; and the fi Keywords: NP1-encoding transcripts were expressed abundantly. In addition, the protein expression pro le of human Human bocavirus bocavirus type 2 (HBoV2) was examined in parallel by transfection of a nearly full-length clone in A549 cells, Gene expression which is similar to that of HBoV1. Moreover, our results showed that, unlike human parvovirus B19 infection, Protein expression of the HBoV1 proteins only does not induce cell cycle arrest and apoptosis of A549 cells. © 2010 Elsevier Inc. All rights reserved. Introduction However, HBoV1 acute infection with high viral loads in respiratory samples (N10,000 copies/ml) and increased IgG or IgM detection have Human Bocavirus (HBoV) is one of the recently identified been frequently associated with acute respiratory illnesses (Allander respiratory viruses, and tentatively is classified in the genus Bocavirus et al., 2005; Kantola et al., 2008), which indicates an apparent within the subfamily of Parvovirinae of the Parvoviridae family etiological link to respiratory illnesses. In addition to respiratory (Cotmore and Tattersall, 2005). Other members in the Bocavirus illnesses, HBoV1 is associated with gastroenteritic diseases (Lee et al., genus are bovine parvovirus type 1 (BPV1) (Chen et al., 1986), minute 2007; Arnold et al., 2006; Lau et al., 2007; Vicente et al., 2007; virus of canines (MVC) (Schwartz et al., 2002). Different species of Albuquerque et al., 2007), a characteristic that shares with two closely HBoV have been identified in humans including the prototype HBoV related animal bocaviruses. The largest genome of HBoV1 that has in respiratory samples as well as HBoV2 and HBoV3 in feces (Kapoor been sequenced is 5299 nts, which lacks both termini; therefore, it is et al., 2009; Arthur et al., 2009). not infectious. To date, both termini of the HBoV genome have not HBoV1 was initially identified from nasopharyngeal aspirates of been sequenced; therefore, an HBoV1 infectious DNA clone has not patients with lower respiratory infections (Allander et al., 2005). The been described. Recently, new species of human Bocavirus, HBoV2 and HBoV1 genome has been frequently detected worldwide, ranging HBoV3, were identified in human stool specimens. HBoV2 has a from 2% to 19% in respiratory specimens from children under 2 years genomic organization identical to that of HBoV1, but the HBoV2 NS1, old with acute respiratory illnesses (Allander et al., 2005; Arden et al., NP1, and VP1 proteins have only 78%, 67%, and 80% identity to those of 2006; Arnold et al., 2006; Bastien et al., 2006; Choi et al., 2006; HBoV1, respectively. Further studies are necessary, however, to Foulongne et al., 2006; Ma et al., 2006; Sloots et al., 2006; Weissbrich identify potential associations of HBoV2 and HBoV3 with clinical et al., 2006; Lin et al., 2007, 2008). HBoV1 is associated with acute symptoms or disease (Kapoor et al., 2009; Arthur et al., 2009). expiratory wheezing and pneumonia (Allander et al., 2007; Kahn, A cell culture system of HBoV1 infection has been recently 2008; Schildgen et al., 2008), and is commonly detected in association established (Dijkman et al., 2009); however, it is inefficient in that with other respiratory viruses (Kahn, 2008; Schildgen et al., 2008). HBoV1 transcripts were only detected by reverse-transcription (RT)- PCR. Six transcripts of HBoV1 were identified from HBoV1-infected differentiated human airway epithelial cells (Dijkman et al., 2009). ⁎ Corresponding author. Department of Microbiology, Molecular Genetics and The abundance of these transcripts and their coding capabilities are Immunology, University of Kansas Medical Center, Mail Stop 3029, 3901 Rainbow not yet understood. Two non-structural proteins NS1 and NP1 were Blvd., Kansas City, KS 66160, USA. Fax: +1 913 588 7295. E-mail address: [email protected] (J. Qiu). predicted, but the NS1 seems to lack the C-terminus compared to the 1 These authors contributed equally. full length NS1 of BPV1 and MVC (Qiu et al., 2007; Sun et al., 2009). 0042-6822/$ – see front matter © 2010 Elsevier Inc. All rights reserved. doi:10.1016/j.virol.2010.04.014 146 A.Y. Chen et al. / Virology 403 (2010) 145–154 The full length sequence of infectious MVC DNA (Genbank accession infection system (Dijkman et al., 2009), with the exception of an extra no.: FJ214110) shows 52.6% identity to HBoV1, while the NS1, NP1 splice site (A1-1) in the first intron. This A1-1 splice site is present in and VP1 proteins of MVC are 38.5%, 39.9% and 43.7% identical to those BPV1 RNA during BPV1 infection (Qiu et al., 2007), but not in MVC RNA of HBoV1, respectively (Sun et al., 2009). We have previously from MVC infection (Sun et al., 2009). determined the transcription profiles of the BPV1 and MVC during To determine the relative abundance of each HBoV1 mRNA, we infection (Qiu et al., 2007; Sun et al., 2009). In BPV1, the left ORF used seven anti-sense HBoV1 probes to protect individual HBoV1 encodes the non-structural protein NS1, at approximately 100 kDa, mRNA. A schematic diagram of the seven anti-sense HBoV1 probes and proteins at relatively small sizes are proposed as NS2 (Qiu et al., with their putative protected bands and nucleotide numbers (nt) is 2007; Lederman et al., 1987). The mid-ORF is thought to encode the shown in Fig. 1A. BPV1 abundant non-structural protein NP1 at 28 kDa, and the right ORF contains the coding sequences for the overlapping capsid protein Probe PD1 genes VP1 and VP2 (Qiu et al., 2007; Lederman et al., 1987). Both the NS1 and NP1 of MVC are important in DNA replication of MVC (Sun Probe PD1, spanning the putative promoter (P3) and the first et al., 2009). The NP1 of HBoV1 and BPV1 can supplement the lack of donor site (D1), protected bands of 133 and 55 nts. These bands function of the MVC NP1 in replication of an NP1 knock-out MVC mapped the RNA initiation site at nt 187 and the first splice donor site infectious clone to some degree (Sun et al., 2009); however, the NP1 (D1) at nt 242. Similar to that of BPV1 and MVC, the first exon of HBoV proteins of the bocaviruses share no similarity to any proteins of the is short, containing only 55 nts (Fig. 1B, lane 2). Approximately 75% of other parvoviruses. Detection of HBoV proteins either during infection HBoV1 RNAs were spliced at the D1 donor site. Multiple bands or in transfection has not been reported. centralized at nt 55 were considered as spliced RNA (Qiu et al., 2002). In this study, we generated a comprehensive transcription profile of HBoV1 by transfecting a replicative chimeric HBoV1 genome in 293 Probe PA1-1 cells and a non-replicative genome in A549 cells. We studied the expression profiles of both the structural and non-structural proteins Probe PA1-1, spanning the first acceptor site of the first intron (A1- of HBoV in detail. Transcripts encoding the left ORF (NS1) are either 1), protected bands of 182 and 120 nts (Fig. 1B, lane 3). The 120-nt spliced or unspliced at a small intron that lies in the middle of the band mapped the first 3′ splice acceptor site at nt 920, which is similar genome. Thus, the spliced transcripts are able to encode a large to the location of the acceptor site in BPV1 RNA. In addition, as seen in nonstructural protein NS1 at approximately 100kDa, which is BPV1, only a small portion of RNAs, less than 10% of spliced RNA at the comparable to the NS1 of MVC and BPV1; while the unspliced D1 donor site, were spliced at the A1-1 acceptor site (Qiu et al., 2007). transcripts encode a relatively small nonstructural protein NS1-70 at approximately 70 kDa. Probe PA1-2 and PA1-2/D2 Results Probe PA1-2, spanning the second acceptor of the first intron (A1-2), protected bands of 162 and 117 nts (Fig. 1C, lane 2). Probe PA1-2/D2, Determine the relative abundance of HBoV1 transcripts by RNase spanning the second acceptor of the first intron (A1-2) and the donor protection assay (RPA) site of the small intron (D2), protected bands of 225, 122 and 168 nts (Fig. 1C, lane 3). These bands protected from both probes confirmed the A transcription map of the HBoV1 has been reported, which was location of the A1-2 acceptor and the D2 donor at nt 2043 and nt 2165, obtained from HBoV1 RNA isolated from HBoV1-infected differentiated respectively.
Load more

Recommended publications
  • Serodiagnosis of Human Bocavirus Infection
    MAJOR ARTICLE Serodiagnosis of Human Bocavirus Infection Kalle Kantola,1 Lea Hedman,1,2 Tobias Allander,4,5 Tuomas Jartti,3 Pasi Lehtinen,3 Olli Ruuskanen,3 Klaus Hedman,1,2 and Maria So¨derlund-Venermo1 1Department of Virology, Haartman Institute, University of Helsinki, and 2Helsinki University Central Hospital Laboratory Division, Helsinki, and 3Department of Pediatrics, Turku University Hospital, Turku, Finland; and 4Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, and 5Department of Clinical Microbiology, Karolinska University Hospital, Stockholm, Sweden (See the editorial commentary by Simmonds on pages 547–9) Background. A new human-pathogenic parvovirus, human bocavirus (HBoV), has recently been discovered and associated with respiratory disease in small children. However, many patients have presented with low viral DNA loads, suggesting HBoV persistence and rendering polymerase chain reaction–based diagnosis problematic. Moreover, nothing is known of HBoV immunity. We examined HBoV-specific systemic B cell responses and assessed their diagnostic use in young children with respiratory disease. Patients and methods. Paired serum samples from 117 children with acute wheezing, previously studied for 16 respiratory viruses, were tested by immunoblot assays using 2 recombinant HBoV capsid antigens: the unique part of virus protein 1 and virus protein 2. Results. Virus protein 2 was superior to the unique part of virus protein 1 with respect to immunoreactivity. According to the virus protein 2 assay, 24 (49%) of 49 children who were positive for HBoV according to polymerase chain reaction had immunoglobulin (Ig) M antibodies, 36 (73%) had IgG antibodies, and 29 (59%) exhibited IgM antibodies and/or an increase in IgG antibody level.
    [Show full text]
  • Investigation of Human Bocavirus in Pediatric Patients with Respiratory Tract Infection
    Original Article Investigation of human bocavirus in pediatric patients with respiratory tract infection Ayfer Bakir1, Nuran Karabulut1, Sema Alacam1, Sevim Mese1, Ayper Somer2, Ali Agacfidan1 1 Department of Medical Microbiology, Division of Virology and Fundamental Immunology, Istanbul Faculty of Medicine, Istanbul University, Istanbul, Turkey 2 Department of Pediatric Infectious Disease, Istanbul University, Istanbul Faculty of Medicine, Istanbul, Turkey Abstract Introduction: Human bocavirus (HBoV) is a linear single-stranded DNA virus belonging to the Parvoviridae family. This study aimed to investigate the incidence of HBoV and co-infections in pediatric patients with symptoms of viral respiratory tract infection. Methodology: This study included 2,310 patients between the ages of 0-18 in whom HBoV and other respiratory tract viral pathogens were analyzed in nasopharyngeal swab specimens. Results: In the pediatric age group, HBoV was found in 4.5% (105/2310) of the patients and higher in children between the ages of 1 and 5. Mixed infection was detected in 43.8% (46/105) of HBoV positive patients (p = 0.10). Mono and mixed infection rates were higher in outpatients than in inpatients (p < 0.05). Respiratory syncytial virus was significantly higher than the other respiratory viral pathogens (p < 0.001). Conclusions: This study is important as it is one of the rare studies performed on the incidence of HBoV in the Marmara region. In pediatric age group, the incidence of HBoV was found 4.5%. The incidence rate of HBoV in this study was similar to those in studies around the world, but close to low rates. The incidence of HBoV was found higher especially among children between the ages of 1-5 in this study.
    [Show full text]
  • First Report of Feline Bocavirus Associated with Severe Enteritis of Cat
    Advance Publication The Journal of Veterinary Medical Science Accepted Date: 8 Feb 2018 J-STAGE Advance Published Date: 20 Feb 2018 1 Note: Virology 2 3 First report of feline bocavirus associated with severe enteritis of cat 4 in Northeast China, 2015 5 6 Chunguo LIU 1), Fei LIU 2), Zhigang LI 4), Liandong QU 1)* and Dafei LIU 1, 3 )* 7 8 9 1) State Key Lab of Veterinary Biotechnology, Harbin Veterinary Research Institute, 10 Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang 150069, China 11 2) Shanghai Hile Bio–Pharmaceutical CO., LTD. Shanghai, 201403, China 12 3) College of Wildlife Resources, Northeast Forestry University, Harbin, Heilongjiang 13 150040, China 14 4) Wendengying Veterinary station, Weihai, Shandong 264413, China 15 16 Correspondence to: 17 Liu, D. and Qu L., State Key Laboratory of Veterinary Biotechnology, Harbin 18 Veterinary Research Institute of CAAS. No. 678 Haping Road, Xiangfang District, 19 Harbin, Heilongjiang, China. 20 E–mail: [email protected] (Liu, D.) and [email protected] (Qu L.). 21 22 Running Title: FIRST REPORT OF FBoV IN CAT IN CHINA 23 1 24 ABSTRACT 25 Feline bocavirus (FBoV) is a newly identified bocavirus of cats in the family 26 Parvoviridae. A novel FBoV HRB2015–LDF was first identified from the cat with 27 severe enteritis in Northeast China, with an overall positive rate of 2.78% (1/36). 28 Phylogenetic and homologous analysis of the complete genome showed that FBoV 29 HRB2015–LDF was clustered into the FBoV branch and closely related to other 30 FBoVs, with 68.7%–97.5% identities.
    [Show full text]
  • Human Bocavirus in Children
    LETTERS 2. Jouanguy E, Altare F, Lamhamedi S, Revy Human Bocavirus ed product size was 354 bp. In each P, Emilie J-F, Levin M, et al. Interferon- experiment, a negative control was γ–receptor deficiency in an infant with fatal in Children bacille Calmette-Guérin infection. N Engl J included, and positive samples were Med. 1996;26:1956–60. To the Editor: Respiratory tract confirmed by analyzing a second 3. Casanova JL, Blanche S, Emile JF, infection is a major cause of illness in sample. Amplification specificity was Jouanguy E, Lamhamedi S, Altare S, et al. verified by sequencing. Idiopathic disseminated bacillus Calmette- children. Despite the availability of Guérin infection: a French national retro- sensitive diagnostic methods, detect- Nine (3.4%) samples were posi- spective study. Pediatrics. 1996;98:774–8. ing infectious agents is difficult in a tive. Comparison of PCR product 4. Roesler J, Kofink B, Wandisch J, Heyden S, substantial proportion of respiratory sequences of these 9 isolates Paul D, Friedrich W, et al. Listeria monocy- (GenBank accession nos. AM109958– togenes and recurrent mycobacterial infec- samples from children with respirato- tions in a child with complete interferon- ry tract disease (1). This fact suggests AM109966) showed minor differ- gamma-receptor (IFNgammaR1) deficien- the existence of currently unknown ences that occurred at 1 to 4 nucleotide cy: mutational analysis and evaluation of respiratory pathogens. positions, and a high level of sequence therapeutic options. Exp Hematol. 1999; identity (99%–100%) was observed 27:1368–74. A new virus has been recently 5. Dorman SE, Uzel G, Roesler J, Bradley J, identified in respiratory samples from with the NP1 sequences of the previ- Bastian J, Billman G, et al.
    [Show full text]
  • Human Bocavirus in Jordan: Prevalence and Clinical Symptoms
    Original Article Singapore Med J 2011; 52(5) : 365 Human bocavirus in Jordan: prevalence and clinical symptoms in hospitalised paediatric patients and molecular virus characterisation AL-Rousan H O, Meqdam M M, Alkhateeb A, Al-Shorman A, Qaisy L M, Al-Moqbel M S ABSTRACT Keywords: bocavirus, Jordan, real-time PCR, Introduction: This study was aimed at sequencing investigating the prevalence of human bocavirus Singapore Med J 2011; 52(5): 365-369 (HBoV) among Jordanian children hospitalised with lower respiratory tract infection (LRTI) as INTRODUCTION well as the clinical feature associated with HBoV Despite the fact that human knowledge of the viruses that infection, the seasonal distribution of HBoV and infect humans is still incomplete, viral infections lay a the DNA sequencing of HBoV positive samples. heavy disease burden on mankind.(1) Unidentified human viruses may exist, causing acute and chronic diseases. Methods: A total of 220 nasopharyngeal aspirates Recent studies have indicated that acute respiratory were collected from children below 13 years of tract infection (ARTI) is a major factor responsible for age who were hospitalised with LRTI in order the hospitalisation of infants and young children, and Department of Applied Biology, to detect the presence of HBoV using real-time that it is also a major cause of morbidity and mortality Jordan University (2) of Science and polymerase chain reaction assay and direct HBoV worldwide. Respiratory tract infection (RTI) is the Technology, sequencing. third most common cause of death in children.(3) PO Box 3030, Irbid-22110, Recently, human bocavirus (HBoV) has been Jordan Results: HBoV was detected in 20 (9.1 percent) identified as a new virus in the respiratory secretions of AL-Rousan HO, MSc patients, whose median age was four (range Graduate Student Swedish children with lower respiratory tract infections 0.8–12) months.
    [Show full text]
  • Phylogenic Analysis of Human Bocavirus Detected in Children With
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Archive Ouverte en Sciences de l'Information et de la Communication Phylogenic analysis of human bocavirus detected in children with acute respiratory infection in Yaounde, Cameroon Sebastien Kenmoe, Marie-Astrid Vernet, Mohamadou Njankouo-Ripa, Véronique Beng Penlap, Astrid Vabret, Richard Njouom To cite this version: Sebastien Kenmoe, Marie-Astrid Vernet, Mohamadou Njankouo-Ripa, Véronique Beng Penlap, Astrid Vabret, et al.. Phylogenic analysis of human bocavirus detected in children with acute respira- tory infection in Yaounde, Cameroon. BMC Research Notes, BioMed Central, 2017, 10, pp.293. 10.1186/s13104-017-2620-y. hal-02366923 HAL Id: hal-02366923 https://hal-normandie-univ.archives-ouvertes.fr/hal-02366923 Submitted on 19 Nov 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution| 4.0 International License Kenmoe et al. BMC Res Notes (2017) 10:293 DOI 10.1186/s13104-017-2620-y BMC Research Notes RESEARCH NOTE Open Access Phylogenic analysis of human bocavirus detected in children with acute respiratory infection in Yaounde, Cameroon Sebastien Kenmoe1,2,3, Marie‑Astrid Vernet1, Mohamadou Njankouo‑Ripa1, Véronique Beng Penlap2, Astrid Vabret3 and Richard Njouom1* Abstract Objective: Human Bocavirus (HBoV) was frst identifed in 2005 and has been shown to be a common cause of respiratory infections and gastroenteritis in children.
    [Show full text]
  • Human Bocavirus in Children EDITORIAL BOARD Co-Editors: Margaret C
    ® CONCISE REVIEWS OF PEDIATRIC INFECTIOUS DISEASES CONTENTS Human Bocavirus in Children EDITORIAL BOARD Co-Editors: Margaret C. Fisher, MD, and Gary D. Overturf, MD Editors for this Issue: John Bradley Board Members Michael Cappello, MD Charles T. Leach, MD Geoffrey A. Weinberg, MD Ellen G. Chadwick, MD Kathleen McGann, MD Leonard Weiner, MD Janet A. Englund, MD Jennifer Read, MD Charles R. Woods, MD Leonard R. Krilov, MD Jeffrey R. Starke, MD Human Bocavirus in Children John C. Arnold, MD Key Words: human bocavirus (HBoV), new lenges that have led to a slow accrual of younger than 5 years, with the highest rates viruses, respiratory infections, gastroenteritis epidemiologic data. Until recently, there was occurring during winter and early spring. (Pediatr Infect Dis J 2010;29: 557–558) no permissive cell line in which to grow the virus, so the description of “acute infections” DISCOVERY has been limited to detection of viral de- CLINICAL SPECTRUM Human bocavirus (HBoV) was first oxyribonucleic acid (DNA). Such descrip- Although the epidemiologic data detected using nonspecific gene sequencing tions are limited because the presence of show that HBoV is a virus that can be fre- of specimens from human subjects with re- DNA could represent acute infections, pro- quently detected in children and to which an spiratory illness.1 Viral, bacterial, and hu- longed shedding of live virus, or inactive immune response is mounted, its role as a man genetic sequences were identified in 2 virus. In addition, there is no universally pathogen has been controversial. Because libraries of specimens collected at different accepted polymerase chain reaction (PCR) HBoV was first recognized in subjects with times of year.
    [Show full text]
  • Original Article Molecular Detection and Phylogenetic Analysis Of
    Original Article Molecular detection and phylogenetic analysis of Kenyan human bocavirus isolates Dennis Misigo1,2, Dufton Mwaengo3, David Mburu4 1 Department of Microbiology and Parasitology, Kenya Methodist University, Meru, Kenya 2 Institute of Tropical and Infectious Diseases, University of Nairobi, Nairobi, Kenya 3 Department of Medical Microbiology, University of Nairobi, Nairobi, Kenya 4 Department of Biochemistry and Biotechnology, Kenyatta University, Nairobi, Kenya Abstract Introduction: The commonly expected causative agents associated with flu-like symptoms in Kenya are the classical viral pathogens identifiable as influenza virus, adenovirus, parainfluenza virus, enteroviruses, respiratory syncytial virus (RSV) and rhinovirus. However, newer agents have been identified globally that present with illnesses clinically indistinguishable from those caused by the classical pathogens; one of them is human bocavirus. Methodology: A total of 384 specimens were analyzed, primarily to determine if the emerging human bocavirus (HBoV) infections exist in Kenya as coinfections with other respiratory viruses and to describe the genotype of the virus in circulation. In brief, viral nucleic acids were extracted from culture supernatants, amplified by PCR, and sequenced. Results: HBoV DNA was amplified from 1.8% of screened specimens. Coinfection with parainfluenza virus, adenovirus, and enterovirus was 2.5%, 2%, and 1.4%, respectively. Multiple coinfections consisting of HBoV plus two other viruses were found in 3% of specimens. Isolation occurred in the months of January, March, April, August, and November. Retrospective review of clinical parameters indicated that all the individuals complained of non-specific symptoms, mainly fever, coughs, nasal stuffiness, runny noses, and vomiting. Phylogenetically, the GenBank deposited sequences of this study’s isolates cluster closely to the reference strain NC_07455 (HBoV1).
    [Show full text]
  • A Comprehensive RNA-Seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium
    viruses Article A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium Wei Zou 1, Min Xiong 2, Xuefeng Deng 1, John F. Engelhardt 3, Ziying Yan 3 and Jianming Qiu 1,* 1 Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USA; [email protected] (W.Z.); [email protected] (X.D.) 2 The Children’s Mercy Hospital, School of Medicine, University of Missouri Kansas City, Kansas City, MO 64108, USA; [email protected] 3 Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA 52242, USA; [email protected] (J.F.E.); [email protected] (Z.Y.) * Correspondence: [email protected]; Tel.: +1-913-588-4329 Received: 11 December 2018; Accepted: 2 January 2019; Published: 7 January 2019 Abstract: Human bocavirus 1 (HBoV1) infects well-differentiated (polarized) human airway epithelium (HAE) cultured at an air-liquid interface (ALI). In the present study, we applied next-generation RNA sequencing to investigate the genome-wide transcription profile of HBoV1, including viral mRNA and small RNA transcripts, in HBoV1-infected HAE cells. We identified novel transcription start and termination sites and confirmed the previously identified splicing events. Importantly, an additional proximal polyadenylation site (pA)p2 and a new distal polyadenylation site (pA)dREH lying on the right-hand hairpin (REH) of the HBoV1 genome were identified in processing viral pre-mRNA. Of note, all viral nonstructural proteins-encoding mRNA transcripts use both the proximal polyadenylation sites [(pA)p1 and (pA)p2] and distal polyadenylation sites [(pA)d1 and (pA)dREH] for termination.
    [Show full text]
  • Human Polyomavirus Type Six in Respiratory Samples From
    Zheng et al. Virology Journal (2015) 12:166 DOI 10.1186/s12985-015-0390-5 RESEARCH Open Access Human polyomavirus type six in respiratory samples from hospitalized children with respiratory tract infections in Beijing, China Wen-zhi Zheng1†, Tian-li Wei2†, Fen-lian Ma1, Wu-mei Yuan1, Qian Zhang1, Ya-xin Zhang2, Hong Cui2* and Li-shu Zheng1* Abstract Background: HPyV6 is a novel human polyomavirus (HPyV), and neither its natural history nor its prevalence in human disease is well known. Therefore, the epidemiology and phylogenetic status of HPyV6 must be systematically characterized. Methods: The VP1 gene of HPyV6 was detected with an established TaqMan real-time PCR from nasopharyngeal aspirate specimens collected from hospitalized children with respiratory tract infections. The HPyV6-positive specimens were screened for other common respiratory viruses with real-time PCR assays. Results: The prevalence of HPyV6 was 1.7 % (15/887), and children ≤ 5 years of age accounted for 80 % (12/15) of cases. All 15 HPyV6-positive patients were coinfected with other respiratory viruses, of which influenza virus A (IFVA) (8/15, 53.3 %) and respiratory syncytial virus (7/15, 46.7 %) were most common. All 15 HPyV6-positive patients were diagnosed with lower respiratory tract infections, and their viral loads ranged from 1.38 to 182.42 copies/μl nasopharyngeal aspirate specimen. The most common symptoms were cough (100 %) and fever (86.7 %). The complete 4926-bp genome (BJ376 strain, GenBank accession number KM387421) was amplified and showed 100 % identity to HPyV6 strain 607a. Conclusions: The prevalence of HPyV6 was 1.7 % in nasopharyngeal aspirate specimens from hospitalized children with respiratory tract infections, as analyzed by real-time PCR.
    [Show full text]
  • Parvovirus Expresses a Small Noncoding RNA That Plays An
    GENOME REPLICATION AND REGULATION OF VIRAL GENE EXPRESSION crossm Parvovirus Expresses a Small Noncoding Downloaded from RNA That Plays an Essential Role in Virus Replication Zekun Wang,a Weiran Shen,a Fang Cheng,a Xuefeng Deng,a John F. Engelhardt,b Ziying Yan,b Jianming Qiua Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, http://jvi.asm.org/ Kansas City, Kansas, USAa; Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USAb ABSTRACT Human bocavirus 1 (HBoV1) belongs to the species Primate bocaparvo- virus of the genus Bocaparvovirus of the Parvoviridae family. HBoV1 causes acute re- Received 7 December 2016 Accepted 20 spiratory tract infections in young children and has a selective tropism for the apical January 2017 Accepted manuscript posted online 25 surface of well-differentiated human airway epithelia (HAE). In this study, we identi- January 2017 fied an additional HBoV1 gene, bocavirus-transcribed small noncoding RNA (BocaSR), Citation Wang Z, Shen W, Cheng F, Deng X, on October 14, 2017 by UNIV OF KANSAS MEDICAL CENTER within the 3= noncoding region (nucleotides [nt] 5199 to 5338) of the viral genome Engelhardt JF, Yan Z, Qiu J. 2017. Parvovirus of positive sense. BocaSR is transcribed by RNA polymerase III (Pol III) from an intra- expresses a small noncoding RNA that plays an essential role in virus replication. J Virol 91: genic promoter at levels similar to that of the capsid protein-coding mRNA and is e02375-16. https://doi.org/10.1128/JVI.02375-16. essential for replication of the viral DNA in both transfected HEK293 and infected Editor Grant McFadden, The Biodesign HAE cells.
    [Show full text]
  • Articles Should Be No More Than 1,200 Words and Need Bocavirus Not Be Divided Into Sections
    Human Bocavirus and Gastroenteritis was similar to that observed in studies that were not limit- This work was partly financed by the “Convenio Diputación ed to specimens that had already tested negative for other Gipúzkoa-Hospital Donostia” and by a grant from the Spanish microorganisms and in which a wide number of agents Ministerio de Sanidad y Consumo CIBER CB06/06. were investigated (4). Adenoviruses have been associated Dr Vicente is a medical microbiologist at the Hospital with infection of the colon and the gut and are a cause of Donostia, Gipúzkoa, Spain. His research focuses on viral respira- severe gastroenteritis in nonindustrialized countries. In this tory infections and meningococcal infection epidemiology. study, coinfection of adenovirus and HBoV was detected in 1 respiratory specimen but these viruses together were not detected in any fecal sample. References HBoV and parvovirus B19 are the only 2 species of 1. Allander T, Tammi MT, Eriksson M, Bjerkner A, Tiveljung-Lindell the Parvoviridae family that have been associated with A, Andersson B. Cloning of a human parvovirus by molecular disease in humans. To date, HBoV has only been detected screening of respiratory tract samples. Proc Natl Acad Sci U S A. in samples from the respiratory tract and has been associ- 2005;102:12891–6. Erratum Proc Natl Acad Sci U S A. 2005; 102:15712. ated with both upper and lower respiratory tract disease in 2. Ma X, Endo R, Ishiguro N, Ebihara T, Ishiko H, Ariga T, et al. infants and young children. The results of our study show Detection of human bocavirus in Japanese children with lower res- that HBoV is also present in the gastrointestinal tract in piratory tract infections.
    [Show full text]